Ammonium derivatives for the treatment of hepatitis C

ABSTRACT

Compounds of Formula I, including pharmaceutically acceptable salts thereof, are set forth, in addition to compositions and methods of using these compounds. The compounds have activity against hepatitis C virus (HCV) and may be useful in treating those infected with HCV.

CROSS REFERENCE TO RELATED APPLICATION

This application claims the priority of U.S. provisional application Ser. No. 61/756,563 filed Jan. 25, 2013 which is herein incorporated by reference.

FIELD OF THE INVENTION

The invention relates to novel compounds of Formula I, including pharmaceutically acceptable salts thereof, which have activity against hepatitis C virus (HCV), and are useful in treating those infected with HCV. The invention also relates to compositions and methods of using these compounds.

BACKGROUND OF THE INVENTION

Hepatitis C virus (HCV) chronically infects an estimated 170 million people worldwide, with 3 to 4 million infected individuals in the United States alone (Boyer, N. and Marcellin, P. J. Hepatology. 2000, 32:98-112; Alter, M. J., et al. Engl. J. Med. 1999, 341:556-562). Prior to the mid 1990s, transfusion with infected blood products was the main route of HCV transmission. Following the introduction of blood screening methods, transmission via injection drug use became the primary risk factor. Chronic infection often leads to the development of severe liver complications, including fibrosis, cirrhosis, and hepatocellular carcinoma. HCV infection is also the leading cause of orthotopic liver transplantation in the United States. The degree to which disease progression is related to viral and cellular factors is not completely understood.

Considerable heterogeneity is found within the nucleotide and encoded amino acid sequence of the HCV genome (Simmonds, P. J. Gen. Virology. 2004, 85:3173-3188). Based on this sequence diversity, six major genotypes and multiple associated subtypes have been described. The genotypes of HCV differ in their worldwide distribution, and the clinical significance of the genetic heterogeneity of HCV remains elusive despite numerous studies of the possible effect of genotypes on pathogenesis and therapy.

Medical treatment for HCV is limited by the lack of a vaccine or approved therapies that specifically target the virus. Currently, patients undergo treatment with a combination of parenterally administered pegylated alpha-interferon and oral ribavirin. Genotype 1 HCV is the most difficult to treat and elimination of the virus (sustained virologic response) is achieved for only approximately 50% of patients (Fried, M. W. et al. N. Engl. J. Med. 2002, 347:975-982; Zeumzem, S. Nature Clinical Practice. 2008, 5:610-622). This poor treatment response, combined with often severe side effects induced by therapy, highlight a need for improved antiviral drugs with better efficacy and safety profiles.

HCV is a member of the Flaviviridae family of viruses with a single-stranded positive-sense RNA genome. Following infection of host cells, the 9.6 Kb genome is translated into a polyprotein precursor of approximately 3,000 amino acids (reviewed in Lindenbach, B. D. and Rice, C. M. Nature. 2005, 436:933-938; Moradpour, D, Penin, F., and Rice, C. M. Nature Reviews. 2007, 5:453-463). Post-translational processing by both cellular and viral proteases results in the generation of at least 10 separate viral proteins. The structural proteins (which by definition are found in mature virions) include core, E1, E2, and possibly p7, and originate from the amino-terminal region of the polyprotein. The core protein assembles into the viral nucleocapsid. The E1 and E2 glycoproteins form heterodimers that are found within the lipid envelope surrounding the viral particles, and mediate host cell receptor binding and entry of the virus into cells. It is unclear if p7 is a structural protein, and its role in replication has yet to be defined. However p7 is believed to form an ion channel in cellular membranes, preventing acidification of intracellular compartments in which virions are assembled, and it has been shown to be essential for viral replication and assembly. The nonstructural proteins NS2, NS3, NS4A, NS4B, NS5A, and NS5B are produced through maturational cleavages of the carboxy-terminal region of the polyprotein. NS2 along with the amino terminus of NS3 form the NS2-3 metalloprotease which cleaves at the NS2-NS3 junction. Additionally, NS2 is involved in assembly and egress of nascent virions. The NS3 protein contains both a serine protease in its amino-terminal region, and a nucleotide-dependent RNA helicase in its carboxy-terminal region. NS3 forms a heterodimer with the NS4A protein, constituting the active protease which mediates cleavages of the polyprotein downstream of NS3, both in cis, at the NS3-NS4A cleavage site, and in trans, for the remaining NS4A-NS4B, NS4B-NS5A, NS5A-NS5B sites. The complex formation of the NS3 protein with NS4A seems necessary to the processing events, enhancing the proteolytic efficiency at all of the sites. The NS3 protein also exhibits nucleoside triphosphatase and RNA helicase activities. The NS4B protein has been shown to be important for localization of HCV proteins into replication complexes in altered membranous structures within the cell. NS5B encodes an RNA-dependent RNA polymerase that is involved in the replication of HCV.

Subgenomic HCV replicons, containing the untranslated regions 5′ and 3′ to the coding sequence fused to the nonstructural proteins or the full-length polyprotein, are competent for translation, viral protein expression, and replication within cultured cells (Lohmann, V. et al. Science. 1999, 285:110-113; Moradpour, D, Penin, F., and Rice, C. M. Nature Reviews. 2007, 5:453-463). The replicon system has proven valuable for the identification of inhibitors targeting the nonstructural proteins associated with these functions. However, only limited subsets of HCV genotypes have been used to generate functional replicons.

Other systems have been used to study the biology of the HCV structural proteins that mediate the entry into host cells. For example, virus-like-particles made in recombinant baculovirus-infected cells with the HCV core, E1 and E2 proteins have also been used to study the function of the HCV E1 and E2 proteins (Barth, H., et al. J. Biol. Chem. 2003, 278:41003-41012). In addition, pseudotyping systems where the E1 and E2 glycoproteins are used to functionally replace the glycoproteins of retroviruses have been developed (Bartosch, B., Dubuisson, J. and Cosset, F.-L. J. Exp. Med. 2003, 197:633-642; Hsu, M. et al. Proc. Natl. Acad. Sci. USA. 2003, 100:7271-7276). These systems yield HCV pseudoparticles that bind to and enter host cells in a manner which is believed to be analogous to the natural virus, thus making them a convenient tool to study the viral entry steps as well as to identify inhibitors block this process.

Recently, a full-length genotype 2a HCV clone, JFH1, was isolated and demonstrated the ability to replicate in vitro. Through repeated passage and adaptation in cell culture increased titers of infectious virus were produced (Lindenbach, B. D., et al. Science. 2005, 309:623-626; Wakita, T. et al. Nature Med. 2005, 11:791-796). In contrast to the HCV replicon or pseudotyping systems, the infectious virus is useful for studying the complete HCV replication cycle, including identifying inhibitors of not only the replication proteins, but those involved in early steps in virus infection (entry and uncoating) and production of progeny viruses (genome packaging, nucleocapsid assembly, virion envelopment and egress).

Triazines have been disclosed. See WO 2009/091388 and US 2009/0286778.

The invention provides technical advantages, for example, the compounds are novel and are effective against hepatitis C. Additionally, the compounds provide advantages for pharmaceutical uses, for example, with regard to one or more of their mechanism of action, binding, inhibition efficacy, target selectivity, solubility, safety profiles, or bioavailability.

SUMMARY OF THE INVENTION

One aspect of the invention is a compound of Formula I, including pharmaceutically acceptable salts thereof:

wherein

-   a, b and c are nitrogen; -   or a and b are nitrogen, while c is —CH; -   or b and c are nitrogen, while a is —CH; -   or a and c are nitrogen, while b is —CH; -   R¹ is selected from alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl,     cycloalkyl, hydroxycycloalkyl, alkoxycycloalkyl, halocycloalkyl,     cycloalkenyl, indanyl, alkylcarbonyl, and benzyl, wherein the benzyl     moiety is substituted with 0-3 substituents selected from halo,     alkyl, cycloalkyl, alkenyl, alkynyl, hydroxyl, cyano, haloalkyl,     alkoxy, and haloalkoxy; -   R² is selected from alkyl, cycloalkyl, ((Ar¹)alkyl, (Ar¹)cycloalkyl,     ((Ar¹)cycloalkyl)alkyl, ((Ar¹)alkyl)cycloalkyl, and     (((Ar¹)alkyl)cycloalkyl)alkyl; -   Ar¹ is phenyl substituted with 0-3 substituents selected from halo,     hydroxy, cyano, alkyl, cycloalkyl, alkenyl, alkynyl, haloalkyl,     alkoxy, and haloalkoxy; -   R³ is hydrogen, alkyl or cycloalkyl; -   R⁴ is selected from the group of hydrogen, halogen, alkyl, alkoxyl,     alkylamino and alkylthio; -   or R² and R⁴ can be connected by a carbon, oxygen, nitrogen or     sulfur atom to form a ring; -   X is selected from the group of O, NR¹¹, CONR¹¹, NR¹¹NR¹² and     CONR¹¹NR¹²; -   R¹¹ is selected from the group of hydrogen, alkyl, hydroxyalkyl,     alkoxyalkyl, haloalkyl, cycloalkyl, hydroxycycloalkyl,     alkoxycycloalkyl, halocycloalkyl, cycloalkenyl, indanyl,     alkylcarbonyl, and benzyl, wherein the benzyl moiety is substituted     with 0-3 substituents selected from halo, alkyl, haloalkyl, alkoxy,     and haloalkoxy; -   R¹² is selected from the group of hydrogen, alkyl, hydroxyalkyl,     alkoxyalkyl, haloalkyl, cycloalkyl, hydroxycycloalkyl,     alkoxycycloalkyl, halocycloalkyl, cycloalkenyl, indanyl,     alkylcarbonyl, and benzyl, wherein the benzyl moiety is substituted     with 0-3 substituents selected from halo, alkyl, haloalkyl, alkoxy,     and haloalkoxy; -   R⁵ is selected from the group of alkyl, cycloalkyl,     (cycloalkyl)alkyl, (alkyl)cycloalkyl, ((alkyl))cycloalkyl)alkyl,     bridged bicycloalkyl, fused bicycloalkyl, and spiro bicycloalkyl,     and is substituted with 0-4 substituents selected from the group of     halo, alkyl, cycloalkyl, hydroxyalkyl, alkoxyalkyl, hydroxy, alkoxy,     benzyloxy, CO₂R¹³, CON(R¹⁴)(R¹⁵), N(R¹⁴)(R¹⁵), tetrahydrofuranyl,     tetrahydropyranyl, and Ar²; -   or R⁵ is hydrogen, N-alkoxycarbonylpiperidinyl, piperidinonyl, or     Ar³; -   R¹³ is selected from the group of hydrogen, alkyl, hydroxyalkyl,     alkoxyalkyl, ((hydroxyalkyl)alkoxy)alkoxy, and     ((alkoxy)alkoxy)alkoxy; -   R¹⁴ is selected from the group of hydrogen, alkyl, cycloalkyl,     alkylcarbonyl, and alkoxycarbonyl; -   R¹⁵ is hydrogen or alkyl; -   or R¹⁴ and R¹⁵ taken together with the nitrogen to which they are     attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, or     morpholinyl, and is substituted with 0-2 substituents selected from     alkyl, alkylcarbonyl, and alkoxycarbonyl; -   Ar² is selected from the group of phenyl, indanyl,     tetrahydronaphthyl, isochromanyl, benzodioxolyl, pyridinyl,     pyrazolyl, imidazolyl, and triazolyl, and is substituted with 0-3     substituents selected from cyano, halo, alkyl, alkyenyl, haloalkyl,     alkoxy, haloalkoxy, N(R¹⁶)(R¹⁷), and alkylCO; -   R¹⁶ is selected from the group of hydrogen, alkyl, cycloalkyl,     alkylcarbonyl, and alkoxycarbonyl; -   R¹⁷ is hydrogen or alkyl; -   or R¹⁶ and R¹⁷ taken together with the nitrogen to which they are     attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, or     morpholinyl, and is substituted with 0-2 substituents selected from     alkyl, alkylcarbonyl, and alkoxycarbonyl; -   Ar³ is selected from the group of phenyl, indanyl, fluorenyl,     biphenyl, terphenyl, pyridinyl, pyrazolyl, isoxazolyl, imidazolyl,     thiazolyl, triazolyl, benzoxazolyl, indolinyl, and dibenzofuranyl,     and is substituted with 0-3 substituents selected from the group of     cyano, halo, alkyl, alkenyl, haloalkyl, cycloalkyl, (CO₂R¹³)alkyl,     (CO₂R¹³)alkenyl, (CON(R¹⁶)(R¹⁷))alkyl, phenyl, hydroxyl, alkoxy,     haloalkoxy, alkylcarbonyl, CO₂R¹³, CON(R¹⁶)(R¹⁷), and PhCONHSO₂; -   or Ar³ is phenyl substituted with 1 substituent selected from     benzyl, tetrazolyloxy, thiazolyl, phenylpyrazolyl,     methyloxadiazolyl, thiadiazolyl, triazolyl, methyltriazolyl,     tetrazolyl, pyridinyl, and dimethoxypyrimdinyl; and -   R⁶ is alkyl or cycloalkyl; -   R⁷ is alkyl or cycloalkyl; -   R⁸ is alkyl or cycloalkyl; -   or R⁷ and R⁸ can be connected by a carbon, oxygen, nitrogen or     sulfur atom to form a ring; -   or R⁶, R⁷ and R⁸ can be connected by a carbon, oxygen, nitrogen or     sulfur atom to form a bicyclic ring; -   or R⁵ and R⁶ can be connected by a carbon, oxygen, nitrogen or     sulfur atom to form a ring; -   or R⁵, R⁶ and R⁷ can be connected by a carbon, oxygen, nitrogen or     sulfur atom to form a bicyclic or tricyclic ring; -   or R⁵, R⁶, R⁷ and R⁸ can be connected by a carbon, oxygen, nitrogen     or sulfur atom to form a tricyclic or tetracyclic ring.

The invention also relates to pharmaceutical compositions comprising a compound of Formula 1, including a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

In addition, the invention provides one or more methods of treating hepatitis C infection comprising administering a therapeutically effective amount of a compound of Formula I to a patient.

Also provided as part of the invention are one or more methods for making the compounds of Formula I.

The present invention is directed to these, as well as other important ends, hereinafter described.

DETAILED DESCRIPTION OF THE EMBODIMENTS

Unless otherwise specifically set forth elsewhere in the application, these terms shall have the following meanings. “H” refers to hydrogen, including its isotopes, such as deuterium. “Halo” means fluoro, chloro, bromo, or iodo. “Alkyl” means a straight or branched alkyl group composed of 1 to 6 carbons. “Alkenyl” means a straight or branched alkyl group composed of 2 to 6 carbons with at least one double bond. “Cycloalkyl” means a monocyclic ring system composed of 3 to 8 carbons. “Alkylene” means a straight or branched divalent alkyl group. “Alkenylene” means a straight or branched divalent alkyl group with at least one double bond. “Cycloalkylene” means a divalent cycloalkane moiety composed of 3 to 7 carbons and includes gem-divalency (for example 1,1-cyclopropanediyl) as well as non-gem-divalency (for example, 1,4-cyclohexanediyl). “Alkylidinyl” means a divalent alkene substituent where the divalency occurs on the same carbon of the alkene. “Hydroxyalkyl,” “alkoxy” and other terms with a substituted alkyl moiety include straight and branched isomers composed of 1 to 6 carbon atoms for the alkyl moiety. “Haloalkyl” and “haloalkoxy” include all halogenated isomers from monohalo substituted alkyl to perhalo substituted alkyl. “Aryl” includes carbocyclic and heterocyclic aromatic substituents. Phenylene is a divalent benzene ring. “1,4-Phenylene” means 1,4-benzenediyl with respect to regiochemistry for the divalent moiety. Parenthetic and multiparenthetic terms are intended to clarify bonding relationships to those skilled in the art. For example, a term such as ((R)alkyl) means an alkyl substituent further substituted with the substituent R.

The substituents described above may be attached at any suitable point of attachment unless otherwise specified. However, it is understood that the compounds encompassed by the present invention are those that are chemically stable as understood by those skilled in the art. Additionally, the compounds encompassed by the present disclosure are those that are suitably stable for use as a pharmaceutical agent.

The invention includes all pharmaceutically acceptable salt forms of the compounds. Pharmaceutically acceptable salts are those in which the counter ions do not contribute significantly to the physiological activity or toxicity of the compounds and as such function as pharmacological equivalents. These salts can be made according to common organic techniques employing commercially available reagents. Some anionic salt forms include acetate, acistrate, besylate, bromide, camsylate, chloride, citrate, fumarate, glucouronate, hydrobromide, hydrochloride, hydroiodide, iodide, lactate, maleate, mesylate, nitrate, pamoate, phosphate, succinate, sulfate, tartrate, tosylate, and xinofoate. Some cationic salt forms include ammonium, aluminum, benzathine, bismuth, calcium, choline, diethylamine, diethanolamine, lithium, magnesium, meglumine, 4-phenylcyclohexylamine, piperazine, potassium, sodium, tromethamine, and zinc.

Some of the compounds of the invention possess asymmetric carbon atoms (see, for example, the structures below). The invention includes all stereoisomeric forms, including enantiomers and diastereomers as well as mixtures of stereoisomers such as racemates. Some stereoisomers can be made using methods known in the art. Stereoisomeric mixtures of the compounds and related intermediates can be separated into individual isomers according to methods commonly known in the art. The use of wedges or hashes in the depictions of molecular structures in the following schemes and tables is intended only to indicate relative stereochemistry, and should not be interpreted as implying absolute stereochemical assignments.

The invention is intended to include all isotopes of atoms occurring in the present compounds. Isotopes include those atoms having the same atomic number but different mass numbers. By way of general example and without limitation, isotopes of hydrogen include deuterium and tritium. Isotopes of carbon include ¹³C and ¹⁴C. Isotopically-labeled compounds of the invention can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described herein, using an appropriate isotopically-labeled reagent in place of the non-labeled reagent otherwise employed. Such compounds may have a variety of potential uses, for example as standards and reagents in determining biological activity. In the case of stable isotopes, such compounds may have the potential to favorably modify biological, pharmacological, or pharmacokinetic properties.

As set forth above, the invention is directed to compounds of Formula I, including pharmaceutically acceptable salts thereof:

wherein

-   a, b and c are nitrogen; -   or a and b are nitrogen, while c is —CH; -   or b and c are nitrogen, while a is —CH; -   or a and c are nitrogen, while b is —CH; -   R¹ is selected from alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl,     cycloalkyl, hydroxycycloalkyl, alkoxycycloalkyl, halocycloalkyl,     cycloalkenyl, indanyl, alkylcarbonyl, and benzyl, wherein the benzyl     moiety is substituted with 0-3 substituents selected from halo,     alkyl, cycloalkyl, alkenyl, alkynyl, hydroxyl, cyano, haloalkyl,     alkoxy, and haloalkoxy; -   R² is selected from alkyl, cycloalkyl, ((Ar¹)alkyl, (Ar¹)cycloalkyl,     ((Ar¹)cycloalkyl)alkyl, ((Ar¹)alkyl)cycloalkyl, and     (((Ar¹)alkyl)cycloalkyl)alkyl; -   Ar¹ is phenyl substituted with 0-3 substituents selected from halo,     hydroxy, cyano, alkyl, cycloalkyl, alkenyl, alkynyl, haloalkyl,     alkoxy, and haloalkoxy; -   R³ is hydrogen, alkyl or cycloalkyl; -   R⁴ is selected from the group of hydrogen, halogen, alkyl, alkoxyl,     alkylamino and alkylthio; -   or R² and R⁴ can be connected by a carbon, oxygen, nitrogen or     sulfur atom to form a ring; -   X is selected from the group of O, NR¹¹, CONR¹¹, NR¹¹NR¹² and     CONR¹¹NR¹²; -   R¹¹ is selected from the group of hydrogen, alkyl, hydroxyalkyl,     alkoxyalkyl, haloalkyl, cycloalkyl, hydroxycycloalkyl,     alkoxycycloalkyl, halocycloalkyl, cycloalkenyl, indanyl,     alkylcarbonyl, and benzyl, wherein the benzyl moiety is substituted     with 0-3 substituents selected from halo, alkyl, haloalkyl, alkoxy,     and haloalkoxy; -   R¹² is selected from the group of hydrogen, alkyl, hydroxyalkyl,     alkoxyalkyl, haloalkyl, cycloalkyl, hydroxycycloalkyl,     alkoxycycloalkyl, halocycloalkyl, cycloalkenyl, indanyl,     alkylcarbonyl, and benzyl, wherein the benzyl moiety is substituted     with 0-3 substituents selected from halo, alkyl, haloalkyl, alkoxy,     and haloalkoxy; -   R⁵ is selected from the group of alkyl, cycloalkyl,     (cycloalkyl)alkyl, (alkyl)cycloalkyl, ((alkyl))cycloalkyl)alkyl,     bridged bicycloalkyl, fused bicycloalkyl, and spiro bicycloalkyl,     and is substituted with 0-4 substituents selected from the group of     halo, alkyl, cycloalkyl, hydroxyalkyl, alkoxyalkyl, hydroxy, alkoxy,     benzyloxy, CO₂R¹³, CON(R¹⁴)(R¹⁵), N(R¹⁴)(R¹⁵), tetrahydrofuranyl,     tetrahydropyranyl, and Ar²; -   or R⁵ is hydrogen, N-alkoxycarbonylpiperidinyl, piperidinonyl, or     Ar³; -   R¹³ is selected from the group of hydrogen, alkyl, hydroxyalkyl,     alkoxyalkyl, ((hydroxyalkyl)alkoxy)alkoxy, and     ((alkoxy)alkoxy)alkoxy; -   R¹⁴ is selected from the group of hydrogen, alkyl, cycloalkyl,     alkylcarbonyl, and alkoxycarbonyl; -   R¹⁵ is hydrogen or alkyl; -   or R¹⁴ and R¹⁵ taken together with the nitrogen to which they are     attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, or     morpholinyl, and is substituted with 0-2 substituents selected from     alkyl, alkylcarbonyl, and alkoxycarbonyl;

Ar² is selected from the group of phenyl, indanyl, tetrahydronaphthyl, isochromanyl, benzodioxolyl, pyridinyl, pyrazolyl, imidazolyl, and triazolyl, and is substituted with 0-3 substituents selected from cyano, halo, alkyl, alkyenyl, haloalkyl, alkoxy, haloalkoxy, N(R¹⁶)(R¹⁷), and alkylCO;

-   R¹⁶ is selected from the group of hydrogen, alkyl, cycloalkyl,     alkylcarbonyl, and alkoxycarbonyl; -   R¹⁷ is hydrogen or alkyl; -   or R¹⁶ and R¹⁷ taken together with the nitrogen to which they are     attached is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, or     morpholinyl, and is substituted with 0-2 substituents selected from     alkyl, alkylcarbonyl, and alkoxycarbonyl; -   Ar³ is selected from the group of phenyl, indanyl, fluorenyl,     biphenyl, terphenyl, pyridinyl, pyrazolyl, isoxazolyl, imidazolyl,     thiazolyl, triazolyl, benzoxazolyl, indolinyl, and dibenzofuranyl,     and is substituted with 0-3 substituents selected from the group of     cyano, halo, alkyl, alkenyl, haloalkyl, cycloalkyl, (CO₂R¹³)alkyl,     (CO₂R¹³)alkenyl, (CON(R¹⁶)(R¹⁷))alkyl, phenyl, hydroxyl, alkoxy,     haloalkoxy, alkylcarbonyl, CO₂R¹³, CON(R¹⁶)(R¹⁷), and PhCONHSO₂; -   or Ar³ is phenyl substituted with 1 substituent selected from     benzyl, tetrazolyloxy, thiazolyl, phenylpyrazolyl,     methyloxadiazolyl, thiadiazolyl, triazolyl, methyltriazolyl,     tetrazolyl, pyridinyl, and dimethoxypyrimdinyl; and -   R⁶ is alkyl or cycloalkyl; -   R⁷ is alkyl or cycloalkyl; -   R⁸ is alkyl or cycloalkyl; -   or R⁷ and R⁸ can be connected by a carbon, oxygen, nitrogen or     sulfur atom to form a ring; -   or R⁶, R⁷ and R⁸ can be connected by a carbon, oxygen, nitrogen or     sulfur atom to form a bicyclic ring; -   or R⁵ and R⁶ can be connected by a carbon, oxygen, nitrogen or     sulfur atom to form a ring; -   or R⁵, R⁶ and R⁷ can be connected by a carbon, oxygen, nitrogen or     sulfur atom to form a bicyclic or tricyclic ring;

or R⁵, R⁶, R⁷ and R⁸ can be connected by a carbon, oxygen, nitrogen or sulfur atom to form a tricyclic or tetracyclic ring.

More preferred compounds, including pharmaceutically acceptable salts thereof, are selected from the group of

In addition, other preferred compounds, including pharmaceutically acceptable salts thereof, are selected from the group of

Pharmaceutical Compositions and Methods of Treatment

The compounds demonstrate activity against HCV NS5B and can be useful in treating HCV and HCV infection. Therefore, another aspect of the invention is a composition comprising a compound, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

Another aspect of the invention is a composition further comprising a compound having anti-HCV activity.

Another aspect of the invention is a composition where the compound having anti-HCV activity is an interferon or a ribavirin. Another aspect of the invention is where the interferon is selected from interferon alpha 2B, pegylated interferon alpha, consensus interferon, interferon alpha 2A, interferon lambda, and lymphoblastoid interferon tau.

Another aspect of the invention is a composition where the compound having anti-HCV activity is a cyclosporin. Another aspect of the invention is where the cyclosporin is cyclosporin A.

Another aspect of the invention is a composition where the compound having anti-HCV activity is selected from the group consisting of interleukin 2, interleukin 6, interleukin 12, a compound that enhances the development of a type 1 helper T cell response, interfering RNA, anti-sense RNA, Imiqimod, ribavirin, an inosine 5′-monophospate dehydrogenase inhibitor, amantadine, and rimantadine.

Another aspect of the invention is a composition where the compound having anti-HCV activity is effective to inhibit the function of a target selected from HCV metalloprotease, HCV serine protease, HCV polymerase, HCV helicase, HCV NS4B protein, HCV entry, HCV assembly, HCV egress, HCV NS5A protein, IMPDH, and a nucleoside analog for the treatment of an HCV infection.

Another aspect of the invention is a composition comprising a compound, or a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable carrier, an interferon and ribavirin.

Another aspect of the invention is a method of inhibiting the function of the HCV replicon comprising contacting the HCV replicon with a compound or a pharmaceutically acceptable salt thereof.

Another aspect of the invention is a method of inhibiting the function of the HCV NS5B protein comprising contacting the HCV NSSB protein with a compound or a pharmaceutically acceptable salt thereof.

Another aspect of the invention is a method of treating an HCV infection in a patient comprising administering to the patient a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof. In another embodiment the compound is effective to inhibit the function of the HCV replicon. In another embodiment the compound is effective to inhibit the function of the HCV NSSB protein.

Another aspect of the invention is a method of treating an HCV infection in a patient comprising administering to the patient a therapeutically effective amount of a compound, or a pharmaceutically acceptable salt thereof, in conjunction with (prior to, after, or concurrently) another compound having anti-HCV activity.

Another aspect of the invention is the method where the other compound having anti-HCV activity is an interferon or a ribavirin.

Another aspect of the invention is the method where the interferon is selected from interferon alpha 2B, pegylated interferon alpha, consensus interferon, interferon alpha 2A, interferon lambda, and lymphoblastoid interferon tau.

Another aspect of the invention is the method where the other compound having anti-HCV activity is a cyclosporin.

Another aspect of the invention is the method where the cyclosporin is cyclosporin A.

Another aspect of the invention is the method where the other compound having anti-HCV activity is selected from interleukin 2, interleukin 6, interleukin 12, a compound that enhances the development of a type 1 helper T cell response, interfering RNA, anti-sense RNA, Imiqimod, ribavirin, an inosine 5′-monophospate dehydrogenase inhibitor, amantadine, and rimantadine.

Another aspect of the invention is the method where the other compound having anti-HCV activity is effective to inhibit the function of a target selected from the group consisting of HCV metalloprotease, HCV serine protease, HCV polymerase, HCV helicase, HCV NS4B protein, HCV entry, HCV assembly, HCV egress, HCV NS5A protein, IMPDH, and a nucleoside analog for the treatment of an HCV infection.

Another aspect of the invention is the method where the other compound having anti-HCV activity is effective to inhibit the function of target in the HCV life cycle other than the HCV NS5B protein.

“Therapeutically effective” means the amount of agent required to provide a meaningful patient benefit as understood by practitioners in the field of hepatitis and HCV infection.

“Patient” means a person infected with the HCV virus and suitable for therapy as understood by practitioners in the field of hepatitis and HCV infection.

“Treatment,” “therapy,” “regimen,” “HCV infection,” and related terms are used as understood by practitioners in the field of hepatitis and HCV infection.

The compounds of this invention are generally given as pharmaceutical compositions comprised of a therapeutically effective amount of a compound or its pharmaceutically acceptable salt and a pharmaceutically acceptable carrier and may contain conventional excipients. Pharmaceutically acceptable carriers are those conventionally known carriers having acceptable safety profiles. Compositions encompass all common solid and liquid forms including for example capsules, tablets, lozenges, and powders as well as liquid suspensions, syrups, elixirs, and solutions. Compositions are made using common formulation techniques, and conventional excipients (such as binding and wetting agents) and vehicles (such as water and alcohols) are generally used for compositions. See, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 17th edition, 1985.

Solid compositions are normally formulated in dosage units and compositions providing from about 1 to 1000 mg of the active ingredient per dose are preferred. Some examples of dosages are 1 mg, 10 mg, 100 mg, 250 mg, 500 mg, and 1000 mg. Generally, other agents will be present in a unit range similar to agents of that class used clinically. Typically, this is 0.25-1000 mg/unit.

Liquid compositions are usually in dosage unit ranges. Generally, the liquid composition will be in a unit dosage range of about 1-100 mg/mL. Some examples of dosages are 1 mg/mL, 10 mg/mL, 25 mg/mL, 50 mg/mL, and 100 mg/mL. Generally, other agents will be present in a unit range similar to agents of that class used clinically. Typically, this is 1-100 mg/mL.

The invention encompasses all conventional modes of administration; oral and parenteral methods are preferred. Generally, the dosing regimen will be similar to other agents used clinically. Typically, the daily dose will be about 1-100 mg/kg body weight daily. Generally, more compound is required orally and less parenterally. The specific dosing regimen, however, will be determined by a physician using sound medical judgment.

The invention also encompasses methods where the compound is given in combination therapy. That is, the compound can be used in conjunction with, but separately from, other agents useful in treating hepatitis and HCV infection. In these combination methods, the compound will generally be given in a daily dose of about 1-100 mg/kg body weight daily in conjunction with other agents. The other agents generally will be given in the amounts used therapeutically. The specific dosing regimen, however, will be determined by a physician using sound medical judgment.

Some examples of compounds suitable for compositions and methods are listed in Table 1.

TABLE 1 Type of Physiological Inhibitor Brand Name Class or Target Source Company NIM811 Cyclophilin Novartis Zadaxin Inhibitor Sciclone Suvus Immuno- Bioenvision Actilon modulator Coley (CPG10101) Methylene blue TLR9 agonist Batabulin (T67) Anticancer β-tubulin Tularik Inc., South inhibitor San Francisco, CA ISIS 14803 Antiviral antisense ISIS Pharmaceuticals Inc, Carlsbad, CA/Elan Phamaceuticals Inc., New York, NY Summetrel Antiviral antiviral Endo Pharmaceuticals Holdings Inc., Chadds Ford, PA GS 9132 (ACH- Antiviral HCV Achillion/Gilead 806) Inhibitor Pyrazolopyrimidine Antiviral HCV Arrow Therapeutics compounds and Inhibitors Ltd. salts From WO- 2005047288 May 26, 2005 Levovirin Antiviral IMPDH Ribapharm Inc., inhibitor Costa Mesa, CA Merimepodib Antiviral IMPDH Vertex (VX-497) inhibitor Pharmaceuticals Inc., Cambridge, MA XTL-6865 (XTL- Antiviral monoclonal XTL 002) antibody Biopharmaceuticals Ltd., Rehovot, Isreal Telaprevir Antiviral NS3 serine Vertex (VX-950, LY- protease Pharmaceuticals 570310) inhibitor Inc., Cambridge, MA/Eli Lilly and Co. Inc., Indianapolis, IN HCV-796 Antiviral NS5B Wyeth/Viropharma Replicase Inhibitor NM-283 Antiviral NS5B Idenix/Novartis Replicase Inhibitor GL 59728 Antiviral NS5B Gene Labs/ Replicase Novartis Inhibitor GL-60667 Antiviral NS5B Gene Labs/ Replicase Novartis Inhibitor 2′C MeA Antiviral NS5B Gilead Replicase Inhibitor PSI 6130 Antiviral NS5B Roche Replicase Inhibitor R1626 Antiviral NS5B Roche Replicase Inhibitor 2′C Methyl Antiviral NS5B Merck Replicase adenosine Inhibitor JTK-003 Antiviral RdRp Japan Tobacco Inc., inhibitor Tokyo, Japan Levovirin Antiviral ribavirin ICN Pharmaceuticals, Costa Mesa, CA Ribavirin Antiviral ribavirin Schering-Plough Corporation, Kenilworth, NJ Viramidine Antiviral Ribavirin Ribapharm Inc., Prodrug Costa Mesa, CA Heptazyme Antiviral ribozyme Ribozyme Pharmaceuticals Inc., Boulder, CO BILN-2061 Antiviral serine Boehringer protease Ingelheim Pharma inhibitor KG, Ingelheim, Germany SCH 503034 Antiviral serine Schering Plough protease inhibitor Zadazim Immune Immune SciClone modulator modulator Pharmaceuticals Inc., San Mateo, CA Ceplene Immuno- immune Maxim modulator modulator Pharmaceuticals Inc., San Diego, CA CellCept Immuno- HCV IgG F. Hoffmann-La suppressant immuno- Roche LTD, Basel, suppressant Switzerland Civacir Immuno- HCV IgG Nabi suppressant immuno- Biopharmaceuticals suppressant Inc., Boca Raton, FL Albuferon-α Interferon albumin Human Genome IFN-α2b Sciences Inc., Rockville, MD Infergen A Interferon IFN InterMune alfacon-1 Pharmaceuticals Inc., Brisbane, CA Omega IFN Interferon IFN-ω Intarcia Therapeutics IFN-β and EMZ701 Interferon IFN-β and Transition EMZ701 Therapeutics Inc., Ontario, Canada Rebif Interferon IFN-β1a Serono, Geneva, Switzerland Roferon A Interferon IFN-α2a F. Hoffmann-La Roche LTD, Basel, Switzerland Intron A Interferon IFN-α2b Schering-Plough Corporation, Kenilworth, NJ Intron A and Interferon IFN-α2b/ RegeneRx Zadaxin α1- Biopharma. Inc., thymosm Bethesda, MD/ SciClone Pharmaceuticals Inc, San Mateo, CA Rebetron Interferon IFN-α2b/ Schering-Plough ribavirin Corporation, Kenilworth, NJ Actimmune Interferon INF-γ InterMune Inc., Brisbane, CA Interferon-β Interferon Interferon- Serono β-1a Multiferon Interferon Long Viragen/ lasting Valentis IFN Wellferon Interferon Lympho- GlaxoSmithKline blastoid plc, Uxbridge, UK IFN-αn1 Omniferon Interferon natural Viragen Inc., IFN-α Plantation, FL Pegasys Interferon PEGylated F. Hoffmann-La IFN-α2a Roche LTD, Basel, Switzerland Pegasys and Interferon PEGylated Maxim Ceplene IFN-α2a/ Pharmaceuticals immune Inc., San Diego, CA modulator Pegasys and Interferon PEGylated F. Hoffmann La Ribavirin IFN-α2a/ Roche LTD, Basel, ribavirin Switzerland PEG-Intron Interferon PEGylated Schering-Plough IFN-α2b Corporation, Kenilworth, NJ PEG-Intron/ Interferon PEGylated Scheringng-Plough Ribavirin IFN-α2b/ Corporation, ribavirin Kenilworth, NJ IP-501 Liver antifibrotic Indevus protection Pharmaceuticals Inc., Lexington, MA IDN-6556 Liver caspase Idun protection inhibitor Pharmaceuticals Inc., San Diego, CA ITMN-191 Antiviral serine InterMune (R-7227) protease Pharmaceuticals inhibitor Inc., Brisbane, CA GL-59728 Antiviral NS5B Genelabs Replicase Inhibitor ANA-971 Antiviral TLR-7 Anadys agonist Boceprevir Antiviral serine Schering Plough protease inhibitor TMS-435 Antiviral serine Tibotec BVBA, protease Mechelen, Belgium inhibitor BI 201335 Antiviral serine Boehringer protease Ingelheim Pharma inhibitor KG, Ingelheim, Germany MK-7009 Antiviral serine Merck protease inhibitor PF-00868554 Antiviral replicase Pfizer inhibitor ANA598 Antiviral Non- Anadys Nucleoside Pharmaceuticals, NS5B Inc., San Diego, CA, Polymerase USA Inhibitor IDX375 Antiviral Non- Idenix Nucleoside Pharmaceuticals, Replicase Cambridge, MA, Inhibitor USA BILB 1941 Antiviral NS5B Boehringer Polymerase Ingelheim Canada Inhibitor Ltd R&D, Laval, QC, Canada PSI-7851 Antiviral Nucleoside Pharmasset, Polymerase Princeton, NJ, USA Inhibitor PSI-7977 Antiviral Nucleotide Pharmasset, NS5B Princeton, NJ, USA Polymerase Inhibitor VCH-759 Antiviral NS5B ViroChem Pharma Polymerase Inhibitor VCH-916 Antiviral NS5B ViroChem Pharma Polymerase Inhibitor GS 9190 Antiviral NS5B Gilead Polymerase Inhibitor Peg-interferon Antiviral Interferon ZymoGenetics/ lamda Bristol-Myers Squibb

Synthetic Methods

The compounds may be made by methods available in the art, as well as those described below and including variations within the skill of the art. Some reagents and intermediates are known in the art. Other reagents and intermediates can be made by methods known in the art using readily available materials. The variables (e.g. numbered “R” substituents) used to describe the synthesis of the compounds are intended only to illustrate how to make the compounds and are not to be confused with variables used in the claims or in other sections of the specification. The following methods are for illustrative purposes and are not intended to limit the scope of the invention.

Abbreviations used in the schemes generally follow conventions used in the art. Chemical abbreviations used in the specification and examples are defined as follows: “NaHMDS” for sodium bis(trimethylsilyl)amide; “DMF” for N,N-dimethylformamide; “MeOH” for methanol; “NBS” for N-bromosuccinimide; “Ar” for aryl; “TFA” for trifluoroacetic acid; “LAH” for lithium aluminum hydride; “BOC”, “DMSO” for dimethylsulfoxide; “h” for hours; “rt” for room temperature or retention time (context will dictate); “min” for minutes; “EtOAc” for ethyl acetate; “THF” for tetrahydrofuran; “EDTA” for ethylenediaminetetraacetic acid; “Et₂O” for diethyl ether; “DMAP” for 4-dimethylaminopyridine; “DCE” for 1,2-dichloroethane; “ACN” for acetonitrile; “DME” for 1,2-dimethoxyethane; “HOBt” for 1-hydroxybenzotriazole hydrate; “DIEA” for diisopropylethylamine, “Nf” for CF₃(CF₂)₃SO₂—; and “TMOF” for trimethylorthoformate.

Abbreviations are defined as follows: “1×” for once, “2×” for twice, “3×” for thrice, “° C.” for degrees Celsius, “eq” for equivalent or equivalents, “g” for gram or grams, “mg” for milligram or milligrams, “L” for liter or liters, “mL” for milliliter or milliliters, “μL” for microliter or microliters, “N” for normal, “M” for molar, “mmol” for millimole or millimoles, “min” for minute or minutes, “h” for hour or hours, “rt” for room temperature, “RT” for retention time, “atm” for atmosphere, “psi” for pounds per square inch, “conc.” for concentrate, “sat” or “sat'd” for saturated, “MW” for molecular weight, “mp” for melting point, “ee” for enantiomeric excess, “MS” or “Mass Spec” for mass spectrometry, “ESI” for electrospray ionization mass spectroscopy, “HR” for high resolution, “HRMS” for high resolution mass spectrometry, “LCMS” for liquid chromatography mass spectrometry, “HPLC” for high pressure liquid chromatography, “RP HPLC” for reverse phase HPLC, “TLC” or “tlc” for thin layer chromatography, “NMR” for nuclear magnetic resonance spectroscopy, “¹H” for proton, “δ” for delta, “s” for singlet, “d” for doublet, “t” for triplet, “q” for quartet, “m” for multiplet, “br” for broad, “Hz” for hertz, and “α”, “β”, “R”, “S”, “E”, and “Z” are stereochemical designations familiar to one skilled in the art.

CHEMISTRY EXPERIMENTAL

LC/MS Method (i.e., Compound Identification)

All Liquid Chromatography (LC) data were recorded on a Shimadzu LC-10AS or LC-20AS liquid chromotograph using a SPD-10AV or SPD-20A UV-Vis detector and Mass Spectrometry (MS) data were determined with a Micromass Platform for LC in electrospray mode.

HPLC Method (i.e., Compound Isolation)

Compounds purified by preparative HPLC were diluted in methanol (1.2 mL) and purified using a Shimadzu LC-8A or LC-10A or Dionex APS-3000 or Waters Acquity™ automated preparative HPLC system.

Syntheses of Intermediates:

Preparation of 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoic acid

Step 1: To a solution of 2,4,6-trichloro-1,3,5-triazine (15 g) in THF (300 mL) was added 2,2,2-trifluoroethanol (8.14 g) and Hunig's Base (15.63 mL). The resulting mixture was stirred for 16 hours. After removal of most THF and precipitate through a plug washing with THF, the filtrate was concentrate to give a crude that will be used as it is.

Step 2: To a solution of the product in Step 1 above (10 g) in THF (100 mL) was added tert-butyl 4-aminobenzoate (7.79 g) and Hunig's Base (7.04 mL). The resulting mixture was stirred for 16 h. The precipitate was filtered and washed with Et₂O, dried, then washed with water and dried to give 10.6 g of tert-butyl 4-(4-chloro-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoate as a solid.

tert-butyl 4-(4-chloro-6-(2,2,2-trifluoroethoxy)- 1,3,5-triazin-2-ylamino)benzoate MS (M + H)⁺ Calcd. 405.1 MS (M + H)⁺ Observ. 405.0 LC Condition Solvent A 100% Water-0.1% TFA Solvent B 100% ACN-0.1% TFA Start % B  2 Final % B  98 Gradient Time  1.6 min Stop Time  1.8 min Flow Rate  0.8 mL/min Wavelength 220 Solvent Pair ACN-H₂O-0.1% TFA Column Aquity UPLC BEH C18 1.7 um

Step 3: To a slurry of tert-butyl 4-(4-chloro-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoate (3.6 g) and 1-(4-chlorophenyl)cyclopropanamine (1.49 g) in THF (50 mL) was stirred for 5 hours at 80° C. The precipitate was filtrated through a plug washing with THF to give a crude product that was purified by Biotage eluting with 4/1-hexane/ethyl acetate to give 1.8 g of tert-butyl 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoate as a solid.

tert-butyl 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6- (2,2,2-trifluoroethoxy)-1,3 5-triazin-2-ylamino)benzoate MS (M + H)⁺ Calcd. 536.2 MS (M + H)⁺ Observ. 536.0 LC Condition Solvent A 100% Water-0.1% TFA Solvent B 100% ACN-0.1% TFA Start % B  2 Final % B  98 Gradient Time  1.6 min Stop Time  1.8 min Flow Rate  0.8 mL/min Wavelength 220 Solvent Pair ACN-H₂O-0.1% TFA Column Aquity UPLC BEH C18 1.7 um

Step 4: A solution of above tert-butyl 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoate (4 g) and HCl in dioxane (7.46 ml, 4M) was stirred for 4 hours. Concentration gave 3.58 g of 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoic acid as a solid.

4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2- trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoic acid MS (M + H)⁺ Calcd. 480.1 MS (M + H)⁺ Observ. 480.1 LC Condition Solvent A 100% Water-0.1% TFA Solvent B 100% ACN-0.1% TFA Start % B  2 Final % B  98 Gradient Time  1.6 min Stop Time  1.8 min Flow Rate  0.8 mL/min Wavelength 220 Solvent Pair ACN-H₂O-0.1% TFA Column Aquity UPLC BEH C18 1.7 um

Preparation of 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoyl chloride

A mixture of sulfuryl dichloride (1.68 g) and 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoic acid (0.40 g) in CH₂Cl₂ (4 mL) in sealed tube was heated at 106° C. for 16 hours. All the solvents were removed under vacuum to give crude 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoyl chloride (0.42 g) as yellow solid, which was used in the further reactions without purification.

Preparation of 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy-D2)-1,3,5-triazin-2-ylamino)benzoyl chloride

Step 1: iPr₂NEt (2.65 g) was added into the solution of 2,4,6-trichloro-1,3,5-triazine (3.78 g, 20.48 mmol) and 2,2,2-trifluoroethanol-D2 (2 g) in THF (150 mL). The reaction was stirred at room temperature for 16 hours.

Step 2: iPr₂NEt (2.65 g) and methyl 4-aminobenzoate (3.10 g) were added into the solution in step 1 and the reaction was stirred at room temperature for 16 hours. Solvents were removed under vacuum to give a residue, to which 20 ml of water and 100 ml of EtOAc were added. The resulted mixture was stirred t room temperature for 16 hours. The solid was isolated by filtration and dried under vacuum to give methyl 4-(4-chloro-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoate (7 g).

methyl 4-(4-chloro-6-(2,2,2-trifluoroethoxy-D2)- 1,3,5-triazin-2-ylamino)benzoate MS (M + H)⁺ Calcd. 365.1 MS (M + H)⁺ Observ. 565.1 Retention Time  1.99 min LC Condition Solvent A  90% Water-10% Methanol-0.1% TFA Solvent B  10% Water-90% Methanol-0.1% TFA Start % B  0 Final % B 100 Gradient Time  2 min Flow Rate  5 mL/min Wavelength 220 Solvent Pair Water-Methanol-TFA Column PHENOMENEX-LUNA 4.6 × 50 mm S10

Step 3: iPr₂NEt (0.35 g) was added into the solution of methyl 4-(4-chloro-6-(2,2,2-trifluoroethoxy-D2)-1,3,5-triazin-2-ylamino)benzoate (1 g) and 1-(4-chlorophenyl)cyclopropanamine (0.46 g) in THF (15 mL). The reaction was stirred at 70° C. for 16 hours. Solvents were removed under vacuum to give a residue, to which 100 ml of EtOAc were added. The organic phase was washed with water (2×20 mL) and brine (15 mL), dried over MgSO₄ and concentrated under vacuum to give methyl 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy-D2)-1,3,5-triazin-2-ylamino)benzoate (1.30 g) which was used in the further reaction without purification.

methyl 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2- trifluoroethoxy-D2)-1,3 5-triazin-2-ylamino)benzoate MS (M + H)⁺ Calcd. 496.1 MS (M + H)⁺ Observ. 496.0 Retention Time  2.18 min LC Condition Solvent A  90% Water-10% Methanol-0.1% TFA Solvent B  10% Water-90% Methanol-0.1% TFA Start % B  0 Final % B 100 Gradient Time  2 min Flow Rate  5 mL/min Wavelength 220 Solvent Pair Water-Methanol-TFA Column PHENOMENEX-LUNA 4.6 × 50 mm S10

Step 4: The mixture of K₂CO₃ (0.84 g) and methyl 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy-D2)-1,3,5-triazin-2-ylamino)benzoate (0.75 g) in acetone (10 mL) and water (10 mL) in sealed tube was heated at 80° C. for 16 hours. After cooling, the mixture was added with 1N HCl to adjust pH to 2 when white solid formed. This white solid was isolated via filtration and dried to give 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy-D2)-1,3,5-triazin-2-ylamino)benzoic acid (0.75g) which was used in the next step with purification.

4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy- D2)-1,3,5-triazin-2-ylamino)benzoic acid MS (M + H)⁺ Calcd. 482.1 MS (M + H)⁺ Observ. 482.3 Retention Time  2.55 min LC Condition Solvent A  5% ACN:95% Water:10 mM Ammonium Actetate Solvent B  95% ACN:5% Water:10 mM Ammonium Actetate Start % B  0 Final % B 100 Gradient Time  4 min Flow Rate  0.8 mL/min Wavelength 220 Solvent Pair ACN:Water:Ammonium Actetate Column Phenomenex LUNA C18, 50 × 2, 3 u

Step 5: A mixture of sulfuryl dichloride (0.42 g) and 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy-D2)-1,3,5-triazin-2-ylamino)benzoic acid (0.31 g) in CH₂Cl₂ (2 mL) in sealed tube was heated at 90° C. for 16 hours. All the solvents were removed under vacuum to give crude 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy-D2)-1,3,5-triazin-2-ylamino)benzoyl chloride (0.31g) as yellow solid, which was used in the further reactions without purification.

Syntheses of Claim I:

Synthesis of Compound 1001, 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-N,N,N-trimethylethanaminium

iPr₂NEt (26 mg) was added into the solution of 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoyl chloride (50 mg) and 2-amino-N,N,N-trimethylethanaminium chloride hydrochloride (18 mg) in THF (4 mL). The reaction was stirred at room temperature for 16 hours. Solvents were removed under vacuum to give a residue which was purified by preparative HPLC to give 1001 (12 mg).

1001 MS Calcd. 564.2 MS Observ. 564.3 Retention Time  2.86 min LC Condition Solvent A  5% ACN:95% Water:10 mM Ammonium Actetate Solvent B  95% ACN:5% Water:10 mM Ammonium Actetate Start % B  0 Final % B 100 Gradient Time  4 min Flow Rate  0.8 mL/min Wavelength 220 Solvent Pair ACN:Water:Ammonium Actetate Column Phenomenex LUNA C18, 50 × 2, 3 u

Synthesis of Compound 1002, 2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy-D2)-1,3,5-triazin-2-ylamino)benzamido)-N,N,N-trimethylethanaminium

iPr₂NEt (26 mg) was added into the solution of 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy-D2)-1,3,5-triazin-2-ylamino)benzoyl chloride (50 mg) and 2-amino-N,N,N-trimethylethanaminium chloride hydrochloride (18 mg) in THF (4 mL). The reaction was stirred at room temperature for 16 hours. Solvents were removed under vacuum to give a residue which was purified by preparative HPLC to give 1002 (2.56 mg).

1002 MS Calcd. 566.2 MS Observ. 566.1 Retention Time  1.74 min LC Condition Solvent A  90% Water-10% Methanol-0.1% TFA Solvent B  10% Water-90% Methanol-0.1% TFA Start % B  0 Final % B 100 Gradient Time  2 min Flow Rate  5 mL/min Wavelength 220 Solvent Pair Water-Methanol-TFA Column PHENOMENEX-LUNA 4.6 × 50 mm S10

Synthesis of Compound 1003, 2-(2-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoyl)hydrazinyl)-N,N,N-trimethyl-2-oxoethanaminium

To a solution of 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoic acid (50 mg) and TBTU (33.5 mg) in DMF (2 mL) was added 2-hydrazinyl-N,N,N-trimethyl-2-oxoethanaminium hydrochloride (17.5 mg) and iPr₂NEt (0.073 mL). After stirring at room temperature for 3 hours, the mixture was purified by preparative HPLC to give 1003 (30 mg).

1003 MS Calcd. 593.2 MS Observ. 593.0 Retention Time  2.90 min LC Condition Solvent A  90% Water-10% Methanol-0.1% TFA Solvent B  10% Water-90% Methanol-0.1% TFA Start % B  30 Final % B 100 Gradient Time  4 min Flow Rate  0.8 mL/min Wavelength 220 Solvent Pair Water-Methanol-TFA Column PHENOMENEX-LUNA 2.0 × 30 mm 3 um

Synthesis of Compound 1004, 3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-N,N,N-trimethylpropan-1-aminium

To a solution of 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoic acid (50 mg) and TBTU (33.5 mg) in DMF (2 mL) was added 3-amino-N,N,N-trimethylpropan-1-aminium TFA salt (22.2 mg) and iPr₂NEt (0.073 mL). After stirring at room temperature for 3 hours, the mixture was purified by preparative HPLC to give 1004 (31.5 mg).

1004 MS Calcd. 578.2 MS Observ. 578.1 Retention Time  3.03 min LC Condition Solvent A  90% Water-10% Methanol-0.1% TFA Solvent B  10% Water-90% Methanol-0.1% TFA Start % B  30 Final % B 100 Gradient Time  4 min Flow Rate  0.8 mL/min Wavelength 220 Solvent Pair Water-Methanol-TFA Column PHENOMENEX-LUNA 2.0 × 30 mm 3 um

Synthesis of Compound 1005, (S)-5-carboxy-5-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-N,N,N-trimethylpentan-1-aminium

To a solution of 4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzoic acid (35 mg) and TBTU (25.8 mg) in DMF (2 mL) was added (S)-2-amino-6-(trimethylammonio)hexanoate hydrochloride (16.39 mg) and iPr₂NEt (0.051 mL). After stirring at room temperature for 3 hours, the mixture was purified by preparative HPLC to give 1005 (17 mg).

1005 MS Calcd. 650.2 MS Observ. 650.1 Retention Time  2.01 min LC Condition Solvent A  90% Water-10% Methanol-0.1% TFA Solvent B  10% Water-90% Methanol-0.1% TFA Start % B  30 Final % B 100 Gradient Time  4 min Flow Rate  0.8 mL/min Wavelength 220 Solvent Pair Water-Methanol-TFA Column PHENOMENEX-LUNA 2.0 × 30 mm 3 um

Synthesis of Compound 1006, 3-(4-(4-(1-(4-chlorophenyl)cyclopropylamino)-6-(2,2,2-trifluoroethoxy)-1,3,5-triazin-2-ylamino)benzamido)-N,N,N,2,2-pentamethylpropan-1-aminium

1006 was prepared via the same synthetic procedure for Compound 1005, using 3-amino-N,N,N,2,2-pentamethylpropan-1-aminium as the coupling reagent.

1006 MS Calcd. 606.3 MS Observ. 606.2 Retention Time  3.23 min LC Condition Solvent A  90% Water-10% Methanol-0.1% TFA Solvent B  10% Water-90% Methanol-0.1% TFA Start % B  30 Final % B 100 Gradient Time  4 min Flow Rate  0.8 mL/min Wavelength 220 Solvent Pair Water-Methanol-TFA Column PHENOMENEX-LUNA 2.0 × 50 mm 3 um

Preparation of 2000, 3000 and 4000 have been described in detail in WO-2011/139513:

The following general procedure was applied to synthesize compounds of Claim I:

iPr₂NEt or Et₃N (1-20 eq.) was added into a solution of 2000 (1 eq.), amine (1-1.5 eq.) and TBTU (1-2 eq.) in DMF or THF at room temperature. The reaction was stirred for 16-72 hours at room temperature or increased temperature from 40° C. to 115° C., before quenched with saturated sodium bicarbonate aqueous solution. The aqueous layer was extracted with EtOAc. The combined organic phase was dried over Mg₂SO₄ and concentrated under vacuum to give a crude product, which was purified by preparative HPLC.

2001 Amine Used

Product

MS Calcd. 664.3 MS Observ. 664.2 Retention Time 4.27 min LC Condition Solvent A 90% Water-10% Methanol-0.1% TFA Solvent B 10% Water-90% Methanol-0.1% TFA Start % B 0 Final % B 100 Gradient Time 4 min Flow Rate 0.8 mL/min Wavelength 220 Solvent Pair Water-Methanol-TFA Column PHENOMENEX-LUNA 2.0 × 50 mm 3 um

2002 Amine Used

Product

MS Calcd. 732.4 MS Observ. 732.5 Retention 3.89 min Time LC Condition Solvent A 5:95 methanol:water with 10 mM ammonium acetate Solvent B 95:5 methanol:water with 10 mM ammonium acetate Gradient: 0.5 min hold at 0% B, 0-100% B over 4 minutes, then a 0.5-minute hold at 100% B Flow Rate 0.5 mL/min Wavelength 220 Solvent Pair Water-Methanol-Ammonium Acetate Column Waters BEH C18, 2.0 × 50 mm, 1.7-μm particles

The following general procedure was applied to synthesize compounds of Claim I:

iPr₂NEt or Et₃N (1-20 eq.) was added into a solution of 3000 (1 eq.), amine (1-1.5 eq.) and TBTU (1-2 eq.) in DMF or THF at room temperature. The reaction was stirred for 16-72 hours at room temperature or increased temperature from 40° C. to 115° C., before quenched with saturated sodium bicarbonate aqueous solution. The aqueous layer was extracted with EtOAc. The combined organic phase was dried over Mg₂SO₄ and concentrated under vacuum to give a crude product, which was purified by preparative HPLC.

3001 Amine Used

Product

MS Calcd. 618.3 MS Observ. 618.3 Retention 3.59 min Time LC Condition Solvent A 90% Water-10% Methanol-0.1% TFA Solvent B 10% Water-90% Methanol-0.1% TFA Start % B 0 Final % B 100 Gradient Time 4 min Flow Rate 0.8 mL/min Wavelength 220 Solvent Pair Water-Methanol-TFA Column PHENOMENEX-LUNA 2.0 × 50 mm 3 um

3002 Amine Used

Product

MS Calcd. 632.2 MS Observ. 632.3 Retention Time 3.64 min LC Condition Solvent A 90% Water-10% Methanol-0.1% TFA Solvent B 10% Water-90% Methanol-0.1% TFA Start % B 0 Final % B 100 Gradient Time 4 min Flow Rate 0.8 mL/min Wavelength 220 Solvent Pair Water-Methanol-TFA Column PHENOMENEX-LUNA 2.0 × 50 mm 3 um

3003 Amine Used

Product

MS Calcd. 660.4 MS Observ. 660.2 Retention Time 3.87 min LC Condition Solvent A 90% Water-0% Methanol-0.1% TFA Solvent B 10% Water-90% Methanol-0.1% TFA Start % B 0 Final % B 100 Gradient Time 4 min Flow Rate 0.8 mL/min Wavelength 220 Solvent Pair Water-Methanol-TFA Column PHENOMENEX-LUNA 2.0 × 50 mm 3 um

Preparation of 4001:

Step 1: Compound 4000 (20 mg) and sulfurous dichloride (63 mg) were mixed together. The reaction mixture was heated to 60° C. for 6 hours. Removal of solvents under vacuum provided crude Compound 4000-In which was used in the next step without purification.

Step 2: iPr₂NEt (9.14 mg) was added into a solution of 4000-In from Step 1 and 2-amino-N,N,N-trimethylethanaminium (4.7 mg) in DMF (1 mL) at room temperature. The reaction was stirred for 16 hours at room temperature. Compound 4001 was isolated by preparative HPLC.

4001 MS Calcd. 650.3 MS Observ. 650.2 Retention Time  3.28 min LC Condition Solvent A  90% Water-10% Methanol-0.1% TFA Solvent B  10% Water-90% Methanol-0.1% TFA Start % B  0 Final % B 100 Gradient Time  4 min Flow Rate  0.8 mL/min Wavelength 220 Solvent Pair Water-Methanol-TFA Column PHENOMENEX-LUNA 2.0 × 50 mm 3 um

Biological Methods

Infection assays. HCV pseudoparticles, produced using standardized methodology (Bartosch, B., Dubuisson, J. and Cosset, F.-L. J. Exp. Med. 2003, 197:633-642) were made via a liposome-based transfection procedure of 293T cells with plasmids expressing the murine leukemia virus capsid and polymerase proteins, an MLV genome encoding the luciferase reporter gene, and envelope glycoproteins from either HCV or vesicular stomatitis virus (VSV). The genotype 1a HCV E1 and E2 envelope coding sequences were derived from the H77C isolate (GenBank accession number AF009606). Media containing pseudoparticles was collected 3 days following transfection, filtered, and stored at −20° C. as a viral stock. Infections were performed in 384-well plates by mixing pseudovirus with 1×10⁴ Huh7 cells/well in the presence or absence of test inhibitors, followed by incubation at 37° C. Luciferase activity, reflecting the degree of entry of the pseudoparticles into host cells, was measured 2 days after infection. The specificity of the compounds for inhibiting HCV was determined by evaluating inhibition of VSV pseudoparticle infection.

Compounds and data analysis. Test compounds were serially diluted 3-fold in dimethyl sulfoxide (DMSO) to give a final concentration range in the assay of 50.0 μM to 0.04 pM. Maximum activity (100% of control) and background were derived from control wells containing DMSO but no inhibitor or from uninfected wells, respectively. The individual signals in each of the compound test wells were then divided by the averaged control values after background subtraction and multiplied by 100% to determine percent activity. Assays were performed in duplicate and average EC₅₀ values (reflecting the concentration at which 50% inhibition of virus replication was achieved) were calculated. Compound EC₅₀ data is expressed as A=0.01≤10 nM; B=10-1000 nM. Representative data for compounds are reported in Table 2.

TABLE 2 EC₅₀ EC₅₀ Compd# Structure (1a, nM) (1a, nM) 1001

A 1002

A 1003

64.090 B 1004

A 1005

A 1006

0.115 A 2001

A 3001

A 3002

B 3003

7.512 A 4001

A

It will be evident to one skilled in the art that the present disclosure is not limited to the foregoing illustrative examples, and that it can be embodied in other specific forms without departing from the essential attributes thereof. It is therefore desired that the examples be considered in all respects as illustrative and not restrictive, reference being made to the appended claims, rather than to the foregoing examples, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. 

What is claimed is:
 1. A compound, including a pharmaceutically acceptable salt thereof, which is selected from the group of


2. A compound, including a pharmaceutically acceptable salt thereof, which is selected from the group of


3. A composition comprising a compound of claim 1, and a pharmaceutically acceptable carrier.
 4. A composition comprising a compound of claim 2, and a pharmaceutically acceptable carrier. 